Device for preparing a sample

ABSTRACT

The invention provides a device for preparing a fluid sample, including but not limited to a sample comprising genomic DNA.

RELATED APPLICATIONS

This Application claims the benefit under 35 U.S.C. §119(e) of U.S.Provisional Application Ser. No. 61/625,743, entitled “DEVICE FORPREPARING A SAMPLE” filed on Apr. 18, 2012, and U.S. ProvisionalApplication Ser. No. 61/783,601, entitled “DEVICE FOR PREPARING ASAMPLE” filed on Mar. 14, 2013, the entire contents of both of which areincorporated herein by reference.

FIELD OF INVENTION

The present invention is directed to a device for preparing a fluidsample, including but not limited to samples which include genomic DNA.More particularly, aspects of the present invention are directed to adevice with a reaction chamber and a porous membrane.

SUMMARY OF INVENTION

According to one aspect, a device for preparing a sample is provided.The device includes a body having a chamber with an inlet and a membranepositioned in the body. The membrane has a first side and a second side,where the inlet is positioned on the first side of the membrane. Thedevice also includes a plurality of channels optionally coupled to thebottom of the chamber, where the plurality of channels are optionallypositioned on the second side of the membrane. Each of the plurality ofchannels extends outwardly from the membrane, the plurality of channelsincluding at least a first channel and a second channel, where the firstchannel extends outwardly from a central portion of the membrane, andwhere the second channel extends outwardly from a peripheral portion ofthe membrane.

According to another aspect, a device for preparing a sample isprovided. The device includes a body having a chamber with an inlet anda membrane positioned in the body. The membrane has a first side and asecond side, where the inlet is positioned on the first side of themembrane. The membrane includes at least a first zone and a second zone,where the first zone is the central portion of the membrane and thesecond zone is the peripheral portion of the membrane and there is abarrier which separates the first zone of the membrane from the secondzone of the membrane. The device also includes a plurality of channelscoupled to the bottom of the chamber, where the plurality of channelsare positioned on the second side of the membrane, in some embodiments.

The present invention further encompasses methods of making and/or usingone or more of the embodiments described herein.

Other advantages and novel features of the present invention will becomeapparent from the following detailed description of various non-limitingembodiments of the invention when considered in conjunction with theaccompanying Figures. In cases where the present specification and adocument incorporated by reference include conflicting and/orinconsistent disclosure, the present specification shall control. If twoor more documents incorporated by reference include conflicting and/orinconsistent disclosure with respect to each other, then the documenthaving the later effective date shall control.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 is a schematic view of a device for preparing a sample accordingto one embodiment;

FIGS. 2A-2D are schematic views of a plurality of sample preparationsteps that may be performed with the device illustrated in FIG. 1;

FIG. 3 is a perspective view of a portion of a device for preparing asample according to one embodiment;

FIG. 4 is another perspective view of the first portion of the deviceshown in FIG. 3;

FIG. 5 is a detailed perspective view of the device shown in FIGS. 3 and4;

FIG. 6 is a perspective view of a portion of the device for preparing asample according to one embodiment; and

FIG. 7 is another perspective view of the portion of the device shown inFIG. 6.

Non-limiting embodiments of the present invention will be described byway of example with reference to the accompanying Figures, which areschematic and are not intended to be drawn to scale. For purposes ofclarity, not every component is labeled in every figure, nor is everycomponent of each embodiment of the invention shown where illustrationis not necessary to allow those of ordinary skill in the art tounderstand the invention.

DETAILED DESCRIPTION OF INVENTION

The invention in its broadest sense provides devices and methods of usethereof for positioning or manipulating or concentrating agents within afluid, including but not limited to polymers such as genomic DNA.Aspects of the invention allow the agents to be concentrated intorelatively small portions of the fluid. This may provide a higherconcentration of the agent within a portion of the fluid, or decreaselosses as the agent undergoes processing due to a decrease of contactarea between the agent and the membrane.

Certain aspects of the invention relate to using a chamber forpositioning or manipulating an agent, such as genomic DNA. In someaspects, the chamber is minimally comprised of an inlet port, a porousmembrane that allows fluid but not the agent of interest to passthrough, and a plurality of channels positioned on a side of the porousmembrane opposite the inlet port. The chamber may be operated in a firstmode where a fluid containing agents is introduced into the chamberthrough the inlet port and flowed through the porous membrane in thechamber. Fluid may be introduced through one or more of the channels tomove a portion of the fluid towards a peripheral portion of themembrane. The desired agents may then be positioned on the centralportion of the membrane. Flow may be reversed through the inlet port tomove any agents positioned on the membrane out of the chamber in centralstreamlines that exit the chamber through the first fluid port.

The invention is based in part on devices with chambers (referred toherein interchangeably as a “reaction chamber” or a “fluidic chamber”)that may be used to concentrate a fluid sample, which may containvarious agents, to a smaller volume of fluid. Concentrating samples mayprove useful when relatively small volumes are available for analysis.Additionally or alternatively, concentrating a sample may prove usefulin introducing a sample from a macro-scale environment, such as fromwhere a sample may have been collected, to a micro-scale or nano-scaleenvironment, such as where analysis may be performed on the sample. Inone embodiment, the device is configured to isolate, purify, and thenprocess various types of samples, including, but not limited to DNA frommicroorganisms.

FIG. 1 illustrates one embodiment of a device 100 for preparing a fluidsample. The device includes a body 30 with a chamber 10 and an inlet 12.As shown, the inlet 12 may be centrally positioned at the top of thechamber 10 and is configured to move fluids into and/or out of thechamber 10. The device 100 also includes a porous membrane 14 thatallows fluid but not the agents of interest to pass therethrough. Aplurality of channels 16, 18, 20 are coupled to the bottom of thechamber. As illustrated, the inlet 12 is positioned on a first side ofthe membrane 14 and the plurality of channels 16, 18, 20 are positionedon a second side of the membrane, where the first side is opposite thesecond side. As set forth below, the channels 16 may be used to directflow, typically introduced through the inlet 12, in differentdirections.

Embodiments of the chamber may be constructed with differentconfigurations and dimensions, some examples of which are discussedherein. By way of example, the chamber 10 may provide a diffusive flowpathway between the inlet port 12 and the flow region, which, in manyembodiments, may laterally spread the flow of fluid introduced throughthe inlet port to promote even distribution of agents about the porousmembrane.

The chamber 10 may be shaped differently according to variousembodiments. In one illustrative embodiment, the chamber 10 includes adiffuser portion 8 which is typically designed to smoothly widen ordiffuse flow that enters the flow region from the inlet port withoutsubjecting agents to excessive shear forces. As shown in FIG. 1, in oneembodiment, the diffuser portion 8 of the chamber 10 is substantiallyfrustoconical in shape. In one embodiment, the chamber 10 has asymmetric, truncated cone shape with substantially linear sides thatform an angle of about 60 degrees with a line that extends along acentral axis of the inlet port 12. It is to be appreciated, however,that the chamber may include walls that are angled differently, or thatare gently curved instead of being linear, as aspects of the inventionare not limited in this regard. In one embodiment, the chamber 10 has asymmetric, truncated cone shape with substantially linear sides thatform an angle of about 45 degrees with a line that extends along acentral axis of the inlet port 12. According to some embodiments, thechamber may include flat sides, appearing more like a truncated pyramid.Other embodiments may also include asymmetric chambers.

The inlet port 12 is typically positioned in the central portion of thechamber and is configured to direct a flow of fluid orthogonally towardthe porous membrane 14 of the chamber 10. According to otherembodiments, however, the inlet port 12 may be offset to one side of thechamber. Additionally or alternatively, the inlet port may direct fluidflow toward the membrane at an angle, instead of orthogonally. It isalso to be appreciated that embodiments of the chamber may include aplurality of inlet ports positioned about the diffuser portion 8.

The chamber and/or inlet port, when described as being substantiallyopposed to the membrane 14, are understood to be positioned to directfluid to impinge on a surface of the membrane. That is, at least aportion of the fluid flow is directed to intersect with the membrane 14.

The porous membrane 14 (also referred to herein as a substrate or afilter) is typically positioned to receive fluid flow that is introducedto the chamber from the inlet port 12, as shown in FIG. 1, such that afluid sample passing therethrough may be received on the membrane 14.The membrane typically has a threshold size that relates to the porosityof the membrane and that describes the size or molecular weight ofagents or other constituents that are prevented from passingtherethrough. According to some embodiments, the membrane 14 has athreshold size that prevents the passage of cells, of genomic DNA, ofproteins, and the like, although other threshold sizes are possible, asaspects of the invention are not limited in this respect. Some examplesof membranes include ultrafiltration membranes. According to manyembodiments, the membrane may be chosen such that it does not have anaffinity for agents that may be processed in the chamber and thus doesnot prevent the agent from being removed from the chamber.

As set forth in more detail below, the membrane 14 may comprise aremovable filter material that is held by a frit or support body 28, asshown in FIG. 1. Some operating protocols may utilize a membrane 14 withdifferent threshold sizes, or that are constructed differently, and maybenefit from being removable from the chamber. According to someembodiments, the membrane itself is relatively stiff, such that asupport body may not be required.

In one embodiment, the chamber 10 may include a body section 6 thatdefines a wall of the chamber 10 that lies between the membrane 14 andthe diffuser portion 8. As shown in FIG. 1, the body section 6 of thechamber 10 is substantially cylindrical in shape and extends for arelatively short distance between the diffuser portion 8 of the chamber10 and the membrane 14. In other embodiments, the chamber 10 may beshaped differently, or the diffuser portion 8 of the chamber 10 mayextend directly to the membrane 14, such that there is no body section 6at all in the chamber 10.

A plurality of channels 16, 18, 20 are positioned adjacent the membrane14, and as shown in FIG. 1, the channels 16, 18, 20 are positioned on asecond side of the membrane 14 (i.e. on a side of the membrane oppositethe inlet 12) and they each extend outwardly from the membrane 14. Theparticular embodiment illustrated in FIG. 1 includes a first channel 16which extends outwardly from a central portion of the membrane 14, asecond channel 18 which extends outwardly from a peripheral portion ofthe membrane 14, and a third channel which also extends outwardly from aperipheral portion of the membrane 14. In one illustrative embodiment,the first channel 16 extends outwardly from a central portion of themembrane, and the second and third channels 18, 20 extend outwardly fromperipheral portions. As illustrated, the third channel 20 may bepositioned on a peripheral portion of the membrane opposite the secondchannel 18.

The plurality of channels 16, 18, 20 are configured to be connected toan external pump or valve that controls the proportion of flow thatpasses though the channels. Any vacuum (or positive pressure) producedby the external pump, in turn, causes a vacuum (or pressure) in one ormore selected channels 16, 18, 20 to move the fluid sample in thechamber. For example, if a vacuum is applied within the first channel16, fluid within the chamber 10 will move into the first channel 16 andagents will collect along the central portion of the porous membrane 14.If a vacuum is applied within the second channel 18, fluid within thechamber 10 will move into the second channel 18 and agents may collectalong the peripheral portion of the porous membrane 14, and similarly,if a vacuum is applied within the third channel 20, fluid within thechamber 10 will move into the third channel 20 and agents may collectalong the peripheral portion of the porous membrane 14. As set forth inmore detail below, in one embodiment, a vacuum may be applied within thefirst channel 16 to initially move the fluid sample and its agentstoward the central portion of the membrane 14 (i.e. toward the firstzone 40 of the membrane). Thereafter, a vacuum may be applied within thesecond channel 18 and/or the third channel 20 to move undesired agentsand/or debris towards the peripheral portion of the membrane 14 (i.e.toward the second zone 50 of the membrane 14), thus isolating thedesired agents on the central portion of the membrane 14. It iscontemplated that a vacuum may also be applied within the first channel16 at the same time that a vacuum is being applied within the second andthird channels 18, 20. Flow may be reversed through the first channel 16to move the desired agents on the central portion of the membrane out ofthe chamber 10. In one embodiment, when a vacuum is applied within thefirst channel 16, the fluid flows substantially normal or perpendicularto the membrane 14 such that the desired agents in the fluid samplepress against the central zone of the membrane. When a vacuum is appliedwithin the second and/or third channels 18, 20, the fluid may flow witha tangential component toward the peripheral portion of the membrane 14.

In one embodiment, the membrane 14 includes at least a first zone 40 anda second zone 50, where the first zone 40 is the central portion of themembrane 14 and the second zone is the peripheral portion of themembrane 14. In one embodiment, the second zone 50 substantiallysurrounds the first zone 40, and the second zone 50 may be substantiallyannular shaped. Other shapes are also contemplated, and in oneembodiment, there may be a plurality of second zones 50 as the inventionis not necessarily so limited. In one embodiment, the first zone issubstantially circular shaped, although other shapes are alsocontemplated.

As shown in FIG. 1, in one embodiment, there is a barrier 60 whichseparates the first zone 40 of the membrane 14 from the second zone 50of the membrane 14. The barrier 60 is configured to prevent movement ofthe sample through or within the membrane, to help to fluidly isolatethe first zone 40 from the second zone 50. Thus, when a vacuum is drawnin the second zone 50 (i.e. with second and/or third channel 18, 20),the barrier 60 may be configured to prevent movement of the samplealready positioned on the first zone 40 of the membrane 14.

It is recognized that the barrier 60 could be formed in a variety ofdifferent ways. For example, in one embodiment, the barrier 60 may beformed by a weld on the membrane material. The first and second zones40, 50 of the membrane 14 may be made of one continuous membranematerial with a weld formed therein to isolate the first zone 40 fromthe second zone 50. In another embodiment, the first and second zones40, 50 may be formed of at least two membrane materials and another typeof barrier 60, such as, but not limited to added layers of the membranematerial, or other types of objects which physically separate the twozones 40, 50 may be employed.

The size and shape of the membrane 14 and the barrier 60 may vary, butas shown in FIG. 6, in one embodiment, the membrane 14 includes asubstantially annular shaped portion and the barrier is substantiallyannular in shape. It is also contemplated that the barrier 60 may beformed from an annular washer-like component. It is further contemplatedthat the barrier may be formed with a sealant.

As shown in FIG. 1, in one illustrative embodiment, the body 30 furtherincludes a collar 70 extending upwardly from the chamber inlet 12. Thecollar 70 has a passageway 72 extending therethrough, and the passageway72 has an inlet 74 configured to receive the fluid sample. In oneillustrative embodiment, the collar inlet 74 is larger than the chamberinlet 12. Such a configuration enables the fluid sample to be moreeasily dispensed into the device 100, while also preventing disturbanceto the reaction chamber 10. For example, the collar inlet 74 may beconfigured to receive a robotic probe which is configured to dispense afluid sample into the device 100. In one illustrative embodiment, atleast a portion of the collar passageway is frustoconical in shape as itnarrows from the collar inlet 74 to the chamber inlet 12. However, othershapes are also contemplated as the present invention is not necessarilylimited in this respect. In one embodiment, the collar passageway 72 isconfigured to act as a reservoir to hold a fluid, such as a buffer. Asset forth below, the passageway 72 may be sized and shaped to hold avolume of the fluid sufficient to perform a particular function. Forexample, in one embodiment, the reservoir passageway 72 is configured tohold a volume that is at least approximately five times larger than thevolume of the chamber 10. In one embodiment, the collar 70 includes arestriction 76 which separates the larger reservoir portion of thepassageway 72 from the chamber inlet 12. This restriction 76 isolatesthe chamber 10 such that the reservoir fluid can be replaced withoutdisturbing the chamber 10.

In one embodiment, the device 100 may also be equipped with features toregulate temperature in the chamber 10. According to one embodiment, afrit 28 that lies below and supports the membrane 14 is made of athermally conductive material, like stainless steel, and may be heatedor cooled by an external source, like a thermoelectric module, toregulate temperature. Additionally or alternately, fluid may passthrough the chamber 10 to cool or heat the chamber. The chamber may alsobe equipped with other devices, like a radiant heater that heats fluidin the chamber through non-contact methods, or like an inline heaterthat heats fluids entering the chamber which, in turn, may help maintainuniform temperature conditions throughout the chamber volume.

Broadly speaking, the plurality of channels 16, 18, 20 are configured toreceive fluid that has passed through the membrane from the flow region.As set forth below, the flow through the various channels 16, 18, 20 canbe varied to control the movement of the fluid sample and the agentscontained within the fluid sample. It is however to be appreciated thatthe channels 16, 18, 20 may be used to accomplish other effects, such asheating and/or cooling of the flow region, as discussed herein.

FIGS. 2A-2D illustrate a plurality of sample preparation steps that maybe performed with the device 100 illustrated in FIG. 1. FIG. 2Aillustrates a washing step in which a buffer is passed through thechamber 10 and that buffer passes through both the first and secondzones 40, 50 of the membrane 14. In particular, as illustrated, a vacuummay be applied within the first channel 16, the second channel 18, andthe third channel 20. The fluid buffer may already be positioned withinthe reservoir portion of the passageway 72.

FIG. 2B illustrates an injecting step in which a fluid sample isinjected into the device 100. In one illustrative embodiment, a roboticprobe 80 is used to inject or dispense the fluid sample into the device100. As illustrated, the device 100 may be filled with the buffer fluidwhen the fluid sample is being injected into the device 100. The type offluid sample may vary, as the invention is not necessarily limited inthis respect, but in one embodiment, the fluid sample contains DNA. Inone embodiment, the sample may include a suspension of cells (such asbut not limited to bacteria, yeast, molds, and/or mycoplasma). In oneembodiment, the sample includes isolated nucleic acids varying in lengthfrom about 0.01 megabases to about 1 megabase in a fluid having a volumebetween about 10 μL to about 100 μL. In another embodiment, the sampleincludes isolated nucleic acids varying in length from about 0.01megabases to about 0.1 megabases, and in another embodiment, the sampleincludes isolated nucleic acids varying in length from about 0.1megabases to about 1 megabase.

In one embodiment, focused flow techniques may be employed during theinjection step. In particular, the buffer fluid surrounding the probe 80is utilized to focus the flow of the sample in the chamber 10. Forexample, a vacuum may be applied within the first channel 16 at a firstflow rate. As mentioned above, this will cause the fluid in the chamberto move toward the central portion of the membrane 14. The fluid sampleis injected into the device at a second flow rate. In one embodiment,the first flow rate is greater than the second flow rate, such that thebuffer surrounding the probe 80 also moves toward the membrane. The flowrate of the buffer toward the membrane is approximately equal to thedifference between the first flow rate and the second flow rate. Thissurrounding sheathed buffer flow may act to focus the flow of the sampletoward the membrane 14 by constraining the sample towards the centralportion of the membrane. In one particular embodiment, the first flowrate is approximately 200 microliters/minute, and the second flow rateis approximately 100 microliters/min, thus the resulting flow rate ofthe surrounding buffer is approximately 100 microliters/min. In anotherembodiment, the first flow rate is approximately 100 microliters/minute,and the second flow rate is approximately 50 microliters/min, thus theresulting flow rate of the surrounding buffer is approximately 50microliters/min.

FIG. 2C illustrates an incubation step where there may be no fluid floweither into or out of the device 100. In one embodiment, the temperatureof the chamber is increased using one of the above-described techniques.For example, the temperature of the chamber 10 may be increased toapproximately 37° C.

Thereafter, another washing step may be performed as shown in FIG. 2A.In this washing step, the undesired cellular debris may be moved intothe second peripheral zone of the membrane and the desired agents in thefluid sample, such as for example, the DNA sample, may be retained onthe central portion of the membrane. As shown in FIG. 2A, a vacuum maybe applied in the first central zone 40 of the membrane 14 to keep thebigger desired agents in the fluid sample on the first central zone 40of the membrane 14. As also shown in FIG. 2A, a vacuum may also beapplied in the peripheral second zone 50 of the membrane 14 to movesmaller undesired components in the fluid sample away from the firstcentral zone 40. As mentioned above, the barrier 60 may be configured toprevent the desired agents in the fluid sample from migrating from thefirst zone into the second zone. The device may be configured such thatthe larger particles/agents remain on the central portion of themembrane, whereas the smaller particles/agents move into the peripheralportion of the membrane.

The steps shown in FIGS. 2A-2C may be repeated one or more times. Forexample, a restriction enzyme may thereafter be injected into thechamber, as shown in FIG. 2B, and then the incubation step shown in FIG.2C may be repeated. In one embodiment, a bacteria sample may beintroduced into the chamber and the bacteria sample may remain on themembrane 14 while undergoing cell lysis, DNA extraction, DNA digestion,and/or DNA labeling.

FIG. 2D illustrates the additional step of ejecting the desired samplefrom the device. As shown, this may be done by reversing the flow offluid and applying a positive pressure through the first channel 16 suchthat the sample that has collected on the central portion of themembrane moves up through the chamber and out of the device. The barrier60 may be configured to prevent the undesired debris, etc. in the secondzone of the membrane from migrating over into the first central zone,thus the undesired debris may remain within the device 100. Once thedesired sample has been removed from the device, the undesired debris,etc. that may have accumulated along the peripheral portion of themembrane may be removed from the device.

FIGS. 3-5 illustrate one embodiment of a first portion 200 of a devicefor preparing a sample which is made from a solid starting material, andFIGS. 6 and 7 illustrates another embodiment of a first portion 300 ofthe device for preparing a sample which is injection molded. FIG. 6illustrates a second portion 310 of the device for preparing a sample.The first portion 200, 300 and the second portions 310, when coupledtogether, may form a device which is substantially equivalent to thedevice 100 described above and shown in FIGS. 1-2. Accordingly, likecomponents have been given identical reference numbers with a prime (′)added.

As shown in FIGS. 3-5, the first portion 200 may include a body 30′ withboth the chamber 10′ and the collar 70′ formed within the body 30′. Inthis embodiment, there is a narrow restriction 76′ that separates thechamber 10′ from a reservoir portion of the collar passageway 72′. Inone embodiment, the restriction 76′ is at least three times the heightof the chamber 10′. In another embodiment, the restriction 76′ is atleast five times the height of the chamber 10′.

As shown in FIGS. 6 and 7, the portion 300 may include a body 310 withthe plurality of channels 16′, 18′, 20′ formed within the body 310, andthe membrane 14′ may be configured to be coupled to the body 310 of theportion 300. This body 310 (which includes the channels 16′, 18′, 20′)may be formed separately from the chamber body. One portion may beconfigured to be disposable and another portion may be configured to bereusable. In this particular illustrative embodiment, the first andsecond portions 300, 310 are configured to form a plurality of separatedevices with chambers 10′ for preparing a sample. In one embodiment,there are four isolated chambers 10′ which are separated with an O-ringseal 202. Other configurations and numbers of chambers 10′ are alsocontemplated, as the present invention is not so limited. For example,it is contemplated that a device with such a configuration may be usedwith a fluid dispensing device that includes a plurality of roboticprobes 80.

As shown best in FIG. 5, the body 30′ may include a plurality ofdownwardly extending feet 210 spaced apart around the perimeter of thechamber 10′. A plurality of passages 220 may be formed between the feet210, and the passages 220 may be configured such that fluid can passfrom the chamber 10′ into the second zone 50 of the membrane 14′.

As shown in FIG. 6, in one illustrative embodiment, one sheet ofmembrane material may form a plurality of membranes 14′ in adjacentchambers 10′. It is also contemplated that multiple membrane sheets mayform the plurality of membranes.

As mentioned above, these first and second portions 300, 310 illustratedin FIGS. 3-7, when coupled together, may form a device which issubstantially equivalent to the device 100 described above and shown inFIGS. 1-2. In one embodiment, the first portion 300 is configured as adisposable component. The first portion 300 may be a one-piece componentin which the reservoir 70, restriction 76, chamber 10 and feet 210 areintegrally formed, and for example, the first portion 200 may be molded.In another embodiment, the second portion 310 is configured as adisposable component.

Fluid flow may be controlled through the chamber during the varioussteps with different configurations of pumps and valves. According tosome embodiments, flow is controlled by a first variable flow rate pumpin fluid communication with the first channel 16 and by a secondvariable flow rate pump that is in fluid communication with the secondand third channels 18, 20. It is to be appreciated, however, that otherarrangements of pumps (either pressure or vacuum) and valves may be usedto control flow through the chamber in various modes of operation, asaspects of the invention are not limited in this respect. Additionally,aspects of the invention are not limited to any one type of pump orvalve.

Embodiments of the chamber may be operated by a controller that receivesinformation for a particular operating protocol and, in turn, controlspumps and/or valves to run the system automatically to complete theprotocol. The term ‘automatically’, as used herein, refers to a systemthat is capable of switching between modes of operation without theintervention of an operator, or to a system that is otherwise capable ofaltering operating conditions, such as flow rates or temperatureswithout manual operator intervention, such as by following a predefinedoperating protocol or by controlling the system to predetermined setpoints. The controller and operating protocol combination may beimplemented in any of numerous ways. For example, in one embodiment, thecontroller and operating protocol combination may be implemented usinghardware, software or a combination thereof. When implemented insoftware, the software code can be executed on, any suitable processoror collection of processors, whether provided in a single computer ordistributed among multiple computers. It should be appreciated that anycomponent or collection of components that perform the functionsdescribed herein can be generically considered as one or morecontrollers that control the functions discussed herein. The one or morecontrollers can be implemented in numerous ways, such as with dedicatedhardware, or with general purpose hardware (e.g., one or moreprocessors) that is programmed using microcode or software to performthe functions recited above. The one or more controllers may be includedin one or more host computers, one or more storage systems, or any othertype of computer that may include one or more storage devices coupled tothe one or more controllers.

In this respect, it should be appreciated that one implementation of theembodiments of the present invention comprises at least onecomputer-readable medium (e.g., a computer memory, a floppy disk, acompact disk, a tape, etc.) encoded with an operating protocol in theform of a computer program (i.e., a plurality of instructions), which,when executed by the controller, performs the herein-discussed functionsof the embodiments of the present invention. The computer-readablemedium can be transportable such that the treatment protocol storedthereon can be loaded onto any computer system resource to implement theaspects of the present invention discussed herein. In addition, itshould be appreciated that the reference to an operating protocol orcontroller which, when executed, performs the herein-discussedfunctions, is not limited to an application program running on a hostcomputer. Rather, the term operating protocol is used herein in ageneric sense to reference any type of computer code (e.g., software ormicrocode) that can be employed to program a processor to implement theherein-discussed aspects of the present invention.

The device may also comprise one or more sensors that receiveinformation from the chamber or channels used to connect the chamber toother portions of the device. Such sensors may receive informationregarding pressure, temperature, flow rates, and the like, in anyportion of the chamber or device. The device may also receiveinformation for detectors that are used to analyze or detect thepresence of an agent in a portion of the device.

It should be appreciated that various embodiments of the presentinvention may be formed with one or more of the above-describedfeatures. The above aspects and features of the invention may beemployed in any suitable combination as the present invention is notlimited in this respect. It should also be appreciated that the drawingsillustrate various components and features which may be incorporatedinto various embodiments of the present invention. For simplification,some of the drawings may illustrate more than one optional feature orcomponent. However, the present invention is not limited to the specificembodiments disclosed in the drawings. It should be recognized that thepresent invention encompasses embodiments which may include only aportion of the components illustrated in any one drawing figure, and/ormay also encompass embodiments combining components illustrated inmultiple different drawing figures.

It should be understood that the foregoing description of variousembodiments of the invention are intended merely to be illustrativethereof and that other embodiments, modifications, and equivalents ofthe invention are within the scope of the invention recited in theclaims appended hereto.

1. A device comprising: a body having a chamber with an inlet; amembrane positioned in the body, the membrane having a first side and asecond side, wherein the inlet is positioned on the first side of themembrane, the membrane comprising at least a first zone and a secondzone; a plurality of channels coupled to the bottom of the chamber,wherein the plurality of channels are positioned on the second side ofthe membrane, wherein each of the plurality of channels extendsoutwardly from the membrane, the plurality of channels including atleast a first channel and a second channel, wherein the first channelextends outwardly from the first zone of the membrane, and wherein thesecond channel extends outwardly from the second zone of the membrane;and a controller that controls: a first flow of fluid into the chamberfrom the inlet and through the first channel to move desired agentstoward the first zone of the membrane; and a second flow of fluid intothe chamber from the inlet and through the second channel to allowundesired agents and/or debris to move toward the second zone of themembrane.
 2. The device of claim 1, wherein the chamber is substantiallyfrustoconical in shape.
 3. The device of claim 1, wherein the bodyfurther comprises a collar extending upwardly from the chamber inlet,the collar having a passageway extending therethough, wherein thepassageway has an inlet configured to receive a sample, wherein thepassageway inlet is larger than the chamber inlet.
 4. (canceled)
 5. Thedevice of claim 1, further comprising a barrier that is configured toseparate the first zone from the second zone.
 6. The device of claim 5,wherein the membrane includes a substantially annular shaped portion andthe barrier is substantially annular in shape.
 7. The device of claim 5,wherein the barrier is formed of a weld on the membrane.
 8. The deviceof claim 1, wherein the plurality of channels further comprises a thirdchannel which extends outwardly from the second zone of the membrane. 9.The device of claim 8, wherein the third channel is positioned on thesecond zone of the membrane opposite the second channel.
 10. The deviceof claim 3, wherein the collar is integrally formed with the chamber.11. The device of claim 1, wherein the plurality of channels are formedinto a second body which is formed separately from the chamber body. 12.The device of claim 1, wherein the chamber body comprises a plurality ofdownwardly extending feet spaced around a perimeter of the chamber, witha plurality of passages formed between the feet configured such thatfluid can pass from the chamber into the second zone of the membrane.13. The device of claim 1 for use in preparing a nucleic acid sample.14. A device comprising: a body comprising a chamber with an inlet; amembrane positioned in the body, the membrane having a first side and asecond side, wherein the inlet is positioned on the first side of themembrane, the membrane comprising at least a first zone and a secondzone; and a barrier configured to separate the first zone from thesecond zone, the first zone configured to receive a first flow of fluidfrom the inlet to move desired agents toward the first zone of themembrane, and the second zone configured to receive a second flow offluid from the inlet to allow undesired agents and/or debris to movetoward the second zone of the membrane.
 15. The device of claim 14,further comprising a plurality of channels coupled to the bottom of thechamber, wherein the plurality of channels are positioned on the secondside of the membrane.
 16. The device of claim 14, wherein the membraneincludes a substantially annular shaped portion and the barrier issubstantially annular in shape.
 17. The device of claim 16, wherein thebarrier is formed of a weld on the membrane.
 18. The device of claim 15,wherein the plurality of channels further comprises a third channelwhich extends outwardly from the second zone of the membrane.
 19. Thedevice of claim 18, wherein the third channel is positioned on thesecond zone of the membrane opposite the second channel.
 20. The deviceof claim 14, wherein the chamber is substantially frustoconical inshape.
 21. The device of claim 14, wherein the body further comprises acollar extending upwardly from the chamber inlet, the collar having apassageway therethough, wherein the passageway has an inlet configuredto receive a sample, wherein the passageway inlet is larger than thechamber inlet.
 22. (canceled)
 23. The device of claim 14 for use inpreparing a nucleic acid sample.
 24. A method comprising: introducinginto the device of claim 1 a nucleic acid comprising sample;manipulating the sample with one or more reagents; and eluting nucleicacid from the device.
 25. (canceled)
 26. The device of claim 14, whereinthe body comprises a plurality of feet spaced around a perimeter of thechamber, with a plurality of passages formed between the feet configuredsuch that fluid can pass from the chamber into the second zone of themembrane.
 27. The device of claim 1, wherein the first zone of themembrane comprises a central portion of the membrane and the second zoneof the membrane comprises a peripheral portion of the membrane.
 28. Thedevice of claim 1, further comprising an O-ring positioned at the bottomof the body.
 29. The device of claim 14, further comprising an O-ringpositioned at the bottom of the body.
 30. The device of claim 15,wherein the plurality of channels includes at least a first channel anda second channel, wherein the first channel extends outwardly from thefirst zone of the membrane, and wherein the second channel extendsoutwardly from the second zone of the membrane.
 31. The device of claim30, wherein the first zone of the membrane comprises a central portionof the membrane and the second zone of the membrane comprises aperipheral portion of the membrane.
 32. A method comprising: providing abody having a chamber with an inlet; providing a membrane positioned inthe body, the membrane having a first side and a second side, whereinthe inlet is positioned on the first side of the membrane, the membranecomprising at least a first zone and a second zone; providing a barrierconfigured to separate the first zone from the second zone; providing aplurality of channels coupled to the bottom of the chamber, wherein theplurality of channels are positioned on the second side of the membrane,wherein each of the plurality of channels extends outwardly from themembrane, the plurality of channels including at least a first channeland a second channel, wherein the first channel extends outwardly fromthe first zone of the membrane, and wherein the second channel extendsoutwardly from the second zone of the membrane; flowing a first flow offluid into the chamber from the inlet and through the first channel tomove desired toward the first zone of the membrane; and flowing a secondflow into the chamber from the inlet and through the second channel toallow undesired agents and/or debris to move toward the second zone ofthe membrane.
 33. The method of claim 32, wherein the first zone of themembrane comprises a central portion of the membrane and the second zoneof the membrane comprises a peripheral portion of the membrane.